Wang S-L, Huang J-A, Liu X-Q
Department of Respiratory Medicine, the First Affiliated Hospital of Soochow University Suzhou, Jiangsu, P.R. China.
Eur Rev Med Pharmacol Sci. 2017 Feb;21(3):600-605.
To explore the mechanism by which KIAA1456 acts on alveolar epithelial cells through lentiviral transfection.
After constructing a KIAA1456 gene vector, 293T cells were co-transfected with lentiviral vectors and after incubation cells were examined by fluorescence microscopy. CCL-149 cells were transfected with LV-KIAA1456 and were examined by fluorescence microscopy. The proliferation capacity of transfected CCL-149 cells was evaluated using flow cytometry. The effect of KIAA1456 overexpression on CCL-149 cells proliferation was studied using the CCK-8 method.
The expression level of KIAA1456 in the LV- KIAA1456 group was significantly higher compared with the LV-Con group and the blank group. Compared with the LV-Con and the blank groups, the proportion of responding cells in G2/M phase showed statistically significant differences. Viable cells had adarker color and higher OD value measured by ELISA. Compared with the control and the blank groups, the growth and proliferation in the CCL-149 transfection group were significantly slower.
KIAA1456 gene inhibited the proliferation of CCL-149 cells by negative regulation of the G2/M cell cycle. We suggest that it can be used as a specific target for the treatment of alveolar epithelium.
通过慢病毒转染探讨KIAA1456作用于肺泡上皮细胞的机制。
构建KIAA1456基因载体后，将慢病毒载体与293T细胞共转染，孵育后通过荧光显微镜检查细胞。用LV-KIAA1456转染CCL-149细胞，并通过荧光显微镜检查。使用流式细胞术评估转染的CCL-149细胞的增殖能力。采用CCK-8法研究KIAA1456过表达对CCL-149细胞增殖的影响。
LV-KIAA1456组中KIAA1456的表达水平显著高于LV-Con组和空白组。与LV-Con组和空白组相比，G2/M期反应细胞的比例显示出统计学上的显著差异。活细胞颜色较深，ELISA检测的OD值较高。与对照组和空白组相比，CCL-149转染组的生长和增殖明显较慢。
KIAA1456基因通过对G2/M细胞周期的负调控抑制CCL-149细胞的增殖。我们认为它可作为治疗肺泡上皮的特异性靶点。
